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Preparation and Characterization of a pH-responsive Polymer that Interacts with Microbial Transglutaminase during Affinity Precipitation
Li, Sipeng1; Chen, Jialing1; Zhang, Xuanjun2; Ding, Zhaoyang1,2; Cao, Xuejun1
2018-02
Source PublicationBIOTECHNOLOGY AND BIOPROCESS ENGINEERING
ISSN1226-8372
Volume23Issue:1Pages:31-38
Abstract

Microbial transglutaminase (MTG) has been widely used in the food and pharmaceuticals industries. In this study, MTG was purified using affinity precipitation with an affinity polymer (P-MMDN-T), which was synthesized using a pH-responsive polymer (PMMDN) coupled with L-thyroxin as an affinity ligand. Interactions between MTG and P-MMDN-T were investigated using turbidimetric titration, zeta potential measurements, and low-field nuclear magnetic resonance (LF-NMR). We found different behaviors, architectures, and phase states of pH-dependent interactions between MTG and P-MMDN-T interactions. Binding energetics between MTG and P-MMDN-T were determined by isothermal titration calorimetry (ITC). The isoelectric point (pI) of the affinity polymer was 4.65 and was recovered with 96.7% efficiency after recycling the polymer three times. The optimal adsorption condition was 0.02 mol/L phosphate buffer (pH 6.0) with 1.0 mol/L NaCl at 30.0A degrees C and a ligand density of 50.0 mu mol/g. The maximum elution recoveries of total MTG were 98.44% (protein) with 92.19% (activity) using 0.02 mol/L pH 10.0 Gly-NaOH as the eluent.

KeywordMicrobial Transglutaminase Ph-responsive Polymer Affinity Precipitation Low-field Nuclear Magnetic Resonance Isothermal Titration Calorimetry
DOI10.1007/s12257-017-0366-y
URLView the original
Indexed BySCI
Language英语
WOS Research AreaBiotechnology & Applied Microbiology
WOS SubjectBiotechnology & Applied Microbiology
WOS IDWOS:000427741600005
PublisherKOREAN SOC BIOTECHNOLOGY & BIOENGINEERING
The Source to ArticleWOS
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Cited Times [WOS]:1   [WOS Record]     [Related Records in WOS]
Document TypeJournal article
CollectionFaculty of Health Sciences
Affiliation1.East China Univ Sci & Technol, Dept Bioengn, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China
2.Univ Macau, Fac Hlth Sci, Macau, Peoples R China
Recommended Citation
GB/T 7714
Li, Sipeng,Chen, Jialing,Zhang, Xuanjun,et al. Preparation and Characterization of a pH-responsive Polymer that Interacts with Microbial Transglutaminase during Affinity Precipitation[J]. BIOTECHNOLOGY AND BIOPROCESS ENGINEERING,2018,23(1):31-38.
APA Li, Sipeng,Chen, Jialing,Zhang, Xuanjun,Ding, Zhaoyang,&Cao, Xuejun.(2018).Preparation and Characterization of a pH-responsive Polymer that Interacts with Microbial Transglutaminase during Affinity Precipitation.BIOTECHNOLOGY AND BIOPROCESS ENGINEERING,23(1),31-38.
MLA Li, Sipeng,et al."Preparation and Characterization of a pH-responsive Polymer that Interacts with Microbial Transglutaminase during Affinity Precipitation".BIOTECHNOLOGY AND BIOPROCESS ENGINEERING 23.1(2018):31-38.
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